Biomedical subjects

S Uchida

Publications and source records attributed to S Uchida.

At least 145 records · Page 8Linked to original sources

PubMed1999

Effects of terfenadine, astemizole and epinastine on electrocardiogram in conscious cynomolgus monkeys.

We examined the effects of non-sedative histamine H1 receptor antagonists on the electrocardiogram (ECG) in conscious cynomolgus monkeys. Terfenadine (3 mg kg(-1) h(-1), i.v.) and astemizole (0.3 and 1 mg kg(-1) h(-1), i.v.) caused significant time-dependent increases in the QT interval and QTc Bazett (QTc). However, normal ECG forms were found during a 60-min infusion of epinastine (3 mg kg(-1) h(-1) i.v.). A higher dose of epinastine (10 mg kg(-1) h(-1), i.v.) increased the QTc and PR interval only 5 min after the start of the infusion. The minimum plasma concentrations of terfenadine, astemizole and epinastine which caused QTc prolongation were 85, 35 and over than 3600 ng/ml, respectively. These drugs did not alter the PQ and QRS intervals and did not cause arrhythmia or atrioventricular block. Our results are consistent with the clinical observation that prolongation of QTc is caused by terfenadine and astemizole but not by epinastine. Thus, measurement of QTc in cynomolgus monkey appears to be a useful approach for evaluating the potential cardiotoxicity of histamine H1 receptor antagonists.

Animals↗

PubMed1999

Isolation and characterization of kidney-specific CLC-K2 chloride channel gene promoter.

CLC-K1 and CLC-K2 are highly homologous kidney-specific chloride channels, but they are expressed in the different nephron segments. To understand the molecular mechanisms of kidney-specific and nephron-segment-specific expression of CLC-K channel genes, the rat ClC-K2 gene promoter was cloned and compared with that of CLC-K1. In the 1.5-kb pair 5'-flanking region of the CLC-K2 gene, no TATA box was identified around the transcriptional start site, and the proximal region (-32 to -68) was characterized by a GA-rich motif that had a significant sequence similarity to that of the previously isolated CLC-K1 gene promoter. In contrast, the distal portion did not have significant sequence similarity to that of CLC-K1. Reporter gene assay and gel-retardation analysis revealed that the GA-rich motif and the binding of a specific protein(s) to this element were indispensable for the basal promoter activity of the CLC-K2 gene. These results suggest that the GA-rich element may have an important role in the promoter activities of the kidney-specific CLC-K1 and -K2 genes, but that the GA-element alone is not sufficient for the strict regulation of nephron-segment-specific expression of CLC-K1 and CLC-K2 genes.

Animals↗

PubMed1999

p38 mitogen-activated protein kinase functionally contributes to chondrogenesis induced by growth/differentiation factor-5 in ATDC5 cells.

Recent studies of intracellular signal transduction mechanisms for the transforming growth factor-beta (TGF-beta) superfamily have focused on Smad proteins, but have paid little attention to mitogen-activated protein (MAP) kinase cascades. Here we demonstrate that growth/differentiation factor-5 (GDF-5), but neither bone morphogenetic protein-2 (BMP-2) nor TGF-beta1, fully promotes the early phase of the chondrogenic response by inducing cellular condensation followed by cartilage nodule formation in a mouse chondrogenic cell line, ATDC5. We investigated which, if any, of the three major types of MAP kinase plays a functional role in the promotion of chondrogenesis induced by GDF-5. GDF-5 induced phosphorylation of p38 MAP kinase and extracellular signal-regulated kinase (ERK) but not that of c-Jun N-terminal kinase (JNK). The phosphorylation of p38 MAP kinase was also induced by BMP-2 and TGF-beta1. An inhibitor of p38 and p38 beta MAP kinase, SB202190, showed complete inhibition of cartilage nodule formation but failed to affect alkaline phosphatase (ALP) activity induced by GDF-5. Expression of the type II collagen gene, a hallmark of chondrogenesis in vertebrates, was also induced by GDF-5 treatment and strongly suppressed by SB202190. On the other hand, although an inhibitor of MAP/ERK kinase, PD98059, inhibited the rapid phosphorylation of ERK by GDF-5, it inhibited neither ALP activity nor cartilage nodule formation induced by GDF-5. These results strongly suggest that the p38 MAP kinase cascade is involved in GDF-5 signaling pathways and that a role of the p38 MAP kinase pathway is necessary over a longer period to promote chondrogenesis in ATDC5 cells.

Alkaline Phosphatase↗

PubMed1999

A comparison of period amplitude analysis and FFT power spectral analysis of all-night human sleep EEG.

Zero-cross and zero-derivative period amplitude analysis (PAA) data were compared with power spectral analysis (PSA) data obtained with the fast Fourier transform in all-night sleep EEG from 10 subjects. Although PAA zero-cross-integrated amplitude showed good agreement with PSA power in 0.3-2 Hz, zero-cross analysis appears relatively ineffective in measuring 2-4 Hz and above waves. However, PAA zero-derivative measures of peak-trough amplitude correlated well with PSA power in 2-4 Hz. Thus, while PAA appears able to measure the entire EEG spectrum, the analytic technique should be changed from zero cross to zero derivative at about 2 Hz in human sleep EEG. PAA and PSA both demonstrate robust and interrelated across-night oscillations in three frequency bands: delta (0.3-4 Hz); sigma (12-16 Hz); and fast beta (20-10 Hz). The frequencies between delta and sigma, and between sigma and fast beta, did not show clear across-night oscillations using either method, and the two methods showed lower epoch-to-epoch agreement in these intermediate bands. The causes of this reduced agreement are not immediately clear, nor is it obvious which method gives more valid results. We believe that the three strongly oscillating frequency bands represent fundamental properties of the human sleep EEG that provide important clues to underlying physiological mechanisms. These mechanisms are more likely to be understood if their dynamic properties are preserved and measured naturalistically rather than being forced into arbitrary sleep stages or procrustean models. Both PAA and PSA can be employed for such naturalistic studies. PSA has the advantages of applying the same analytic method across the EEG spectrum and rests on more fully developed theory. Combined zero-cross and zero-derivative PAA demonstrates EEG oscillations that closely parallel those observed with spectral power, and the PAA measures do not rely on assumptions about the spectral composition of the signal. In addition, both PAA techniques can measure the relative contributions of wave amplitude and incidence to total power: These waveform characteristics represent different biological processes and respond differentially to a wide range of experimental conditions.

Adult↗

PubMed1999

The effects of morphine on supraspinal and propriospinal somatocardiac reflexes in anesthetized rats.

In anesthetized rats, the effects of i.v. injection of morphine on supraspinally- and spinally-mediated tachycardiac reflex responses of heart rate (HR) and cardiac sympathetic nerve reflex activity were examined following electrical stimulation of either a non-segmental hindlimb (tibial) or a segmental (3rd or 4th intercostal, IC3-4) afferent nerve. In central nervous system (CNS)-intact rats, the supraspinally-mediated reflex increase in HR in response to tibial afferent nerve stimulation was augmented by morphine, whereas the increase in response to IC3-4 afferent nerve stimulation was variable. Both the supraspinally-mediated myelinated (A) and unmyelinated (C) reflex discharges in the cardiac sympathetic nerve elicited by tibial afferent nerve stimulation were augmented by morphine. The effects of morphine on A- and C-sympathetic reflex discharges elicited by IC3-4 afferent nerve stimulation varied depending upon whether the discharge was of supraspinal or spinal origin. In spinal rats (spinalized at the cervical level), tibial afferent nerve stimulation did not produce any HR response, whereas IC3-4 afferent nerve stimulation produced a reflex increase in both HR and sympathetic reflex discharges of spinal origin. Furthermore, these spinal HR and sympathetic nerve reflex responses were inhibited by morphine. In conclusion, morphine depressed somatocardiac sympathetic reflexes at the spinal level, but enhanced these reflexes at the supraspinal level, leading to different effects of morphine on somatically-induced HR responses of supraspinal and spinal origins depending upon the segmental levels of afferent nerves stimulated.

Afferent Pathways↗

PubMed1999

Epinastine, a nonsedating histamine H1 receptor antagonist, has a negligible effect on HERG channel.

Terfenadine and astemizole rarely cause cardiac arrhythmias by suppressing the cardiac rapid delayed rectifier K+ channel encoded by the human ether-a-go-go-related gene (HERG). Epinastine, however, has not been reported to have the adverse effect. We have therefore compared the effects of epinastine, terfenadine and astemizole on HERG channels expressed in Xenopus oocytes. Terfenadine and astemizole suppressed the HERG current with IC50 of 431 nM and 69 nM, respectively. In contrast, 100 microM epinastine inhibited the HERG current by only 11+/-2.1%. These results may provide an explanation for the difference in the cardiotoxicity between different nonsedating histamine H1 receptor antagonists.

Animals↗

PubMed1999

Synthesis and Structural Ordering of Gradient-Modulus Star Copolymers.

Gradient-modulus star copolymers were synthesized by crosslinking of polystyrene/isoprene tapered anions with divinylbenzene in benzene. The styrene content in the tapered arm was determined by 1H-nuclear magnetic resonance as a parameter of conversion. The composition of the arm showed the gradient structure and was in good agreement with that calculated by reactivity ratios. The solution properties of gradient-modulus star copolymers were examined by static and dynamic light scatterings. The structural ordering of gradient-modulus star copolymers were investigated by means of small-angle X-ray scattering. Gradient-modulus star copolymers formed a body-centered cubic (BCC) structure near the overlap threshold (C*). Above C*, this structure changed to the mixed lattice of BCC and face-centered cubic (FCC) structures. Copyright 1999 Academic Press.

Journal Article↗

PubMed1999

The effect of electro-acupuncture stimulation on the muscle blood flow of the hindlimb in anesthetized rats.

The effect of electro-acupuncture stimulation (EAS) on blood flow in the muscle biceps femoris (MBF) and on mean arterial pressure (MAP) was investigated in anesthetized, artificially ventilated rats. EAS was applied to a hindpaw for 30 s at intensities of 0.1-10.0 mA and at frequencies of 1-20 Hz, and MBF was measured by laser Doppler flowmetry. EAS at less than 1.0 mA, which excited group II fibers maximally and III fibers partially in a saphenous nerve, had no significant effect on MBF or MAP, although both revealed variable responses. EAS at 1.5 mA, which additionally excited group III fibers almost maximally and was subthreshold for group IV fibers, produced a small but significant increase in MBF and MAP. These responses were further increased at 2.0 mA or more, which was suprathreshold for group IV fibers. The increased response of MBF at 10.0 mA was followed by a small decrease in MBF. EAS at 1.5 mA or more also elicited a decrease in renal blood flow (RBF) and an arterial pressor response. Following severance of the bilateral splanchnic nerves, EAS at 10.0 mA induced only a slight increase in MAP and a decrease in MBF. The decrease in MBF was abolished following further severance of the bilateral lumbar sympathetic trunks (LSTs). In conclusion, EAS to a hindpaw at a stimulus strength sufficient to excite group III and IV afferent fibers, particularly group IV afferent fibers, can produce a reflex decrease in MBF via a reflex activation of muscle sympathetic activity, although this decrease in MBF is overridden by an increase in MBF caused passively by a reflex MAP pressor response elicited by a reflex increase, at least in splanchnic sympathetic activity.

Action Potentials↗

PubMed1999

Arterial tonometry in the measurement of the effects of innocuous mechanical stimulation of the neck on heart rate and blood pressure.

Arterial tonometry was used to continuously monitor the effects on heart rate and blood pressure of a series of innocuous mechanical stimuli applied to the neck in conscious humans. The stimuli used were derived from procedures commonly employed in clinical examination and physical therapy of the neck. In alert subjects, the stimuli used generally caused small and sometimes statistically significant decreases in heart rate, systolic pressure and diastolic pressure. In alert subjects, statistically significant decreases in systolic and diastolic pressure were particularly associated with stimuli which involved full rotation of the neck. In the course of the prolonged series of stimuli, some subjects slept or reported being on the verge of sleep. In these subjects, the same stimuli produced mixed effects on heart rate and increases in systolic and diastolic pressure which were significantly different from the effects obtained in fully alert subjects.

Adult↗

PubMed1999

Molecular cloning of CLC chloride channels in Oreochromis mossambicus and their functional complementation of yeast CLC gene mutant.

We have cloned two members of the CLC chloride channel family (OmCLC-3 and OmCLC-5) from gill cDNA libraries of the euryhaline tilapia Oreochromis mosammbicus. At the amino acid level, OmCLC-3 is 90.5% identical to rat CLC-3 and OmCLC-5 is 79.2% identical to rat CLC-5. Ribonuclease protection assay revealed that OmCLC-5 was mainly expressed in the gill, kidney, and intestine in both freshwater- (FW) and seawater- (SW) adapted tilapia. Although the mRNA of OmCLC-3 was broadly expressed in tissues of FW- and SW-adapted tilapia, the most intense signals were observed in the gill, kidney, intestine, and brain. Injection of OmCLC-3 and OmCLC-5 cRNAs into Xenopus oocytes did not elicit chloride currents, but these clones did functionally complement the gef1 phenotype of YPH250(gef), a yeast strain in which a single CLC channel (GEF1) has been disrupted by homologous recombination. These results clearly indicated that CLC channels closely related to the mammalian CLC-3, -4, and -5 subfamily exist also in tilapia and that OmCLC-3 and OmCLC-5 function as intracellular chloride channels.

Amino Acid Sequence↗

PubMed1999

Uterine contractility and blood flow are reflexively regulated by cutaneous afferent stimulation in anesthetized rats.

The effects of cutaneous mechanical afferent stimulation of various skin areas on uterine contractility and blood flow were examined in anesthetized non-pregnant rats. The contractility of the uterus was measured by the balloon method in the uterus. The uterine blood flow was measured by laser Doppler flowmetry. Noxious pinching stimulation of the perineum for 1 min induced an abrupt contraction of the uterus during stimulation. Pinching of a hindpaw or perineum and innocuous brushing of the perineum for 1 min increased uterine blood flow. Stimulation of other skin areas produced no changes in uterine contractility or blood flow. Most uterine responses were abolished by severance of the pelvic nerves, which innervated the uterus. The activity of pelvic parasympathetic efferent nerves to the uterus increased following perineal pinching. All these cutaneous stimulation-induced responses of uterine contractility, blood flow and pelvic efferent nerve activity still existed, and were even augmented, after acute spinalization. These results indicate that cutaneous mechanical sensory stimulation can regulate uterine contractility and blood flow by a segmental spinal reflex mechanism via uterine parasympathetic efferent nerves.

Anesthesia↗

PubMed1999

Safety and efficacy of autologous platelet transfusion in cardiac surgery: comparison of cryopreservation, blood collection on the day before surgery, and blood collection during surgery.

In a group of 39 patients with ischemic heart and valvular disease (January 1997 to May 1998), three platelet collection methods were compared in terms of safety and effectiveness. The methods were: (i) collection of autologous platelets over several weeks and freezing them for storage until surgery (frozen group, 12 patients); (ii) collection of autologous platelets on the day before surgery and preserving them without freezing (fresh group, 8 patients); and (iii) collection of autologous platelets intraoperatively (intraoperative group, 9 patients). Ten patients served as controls (control group). Blood pressure was not significantly affected by platelet collection in the frozen and fresh groups, but both systolic (P < 0.01) and diastolic blood pressure (P < 0.05) decreased significantly after collecting platelets in the intraoperative group. Similarly, heart rate was unaffected by platelet collection in the frozen and fresh groups, while it increased significantly in the intraoperative group (P < 0.05). Blood loss after 24 h was significantly smaller in the fresh group than in the frozen group (P < 0.05). Total blood transfusion volume was significantly smaller in the frozen and fresh groups than in the intraoperative and control groups (P < 0.05). Bleeding time 2 h postoperatively, when administration of autologous platelets had been completed, was reduced compared with immediately postoperative values in all three groups receiving autologous platelets (P < 0.05). However, only the frozen and fresh groups showed a significantly shorter bleeding time than the control group (P < 0.05). In all three groups receiving autologous platelets, the platelet count was significantly increased after administration of autologous platelets, but only the fresh group had a platelet count that was significantly greater than the control group (P < 0.05). From these results we conclude that the frozen and fresh groups received safer treatment than the intraoperative group. Although hemostasis improved after all three regimes of autologous platelet transfusion, only the frozen and fresh groups had a reduced need for allogeneic blood transfusion compared with the control group. For this reason we conclude that the frozen and fresh groups were also superior to the intraoperative group in terms of effectiveness. However, the recovery of platelets after frozen storage was low, and to obtain a good effect with the freezing method it is necessary to collect and store large volumes of platelets. In terms of simplicity, safety, and efficacy, the fresh method seems to be the preferred technique.

Aged↗

PubMed1999

A case of acute pulmonary embolism and acute myocardial infarction with suspected paradoxical embolism after laparoscopic surgery.

We describe the case of a 59-year-old Japanese man who had an acute pulmonary embolism in addition to acute myocardial infarction after a laparoscopic cholecystectomy. The posterior descending coronary artery was totally occluded. and direct percutaneous transluminal balloon angioplasty was performed. The pulmonary embolism was diagnosed by lung perfusion scanning and was treated with anticoagulant therapy. A patent foramen ovale and right-to-left atrial shunting of blood were detected by contrast transesophageal echocardiography. Paradoxical embolism is a rare complication of pulmonary embolism and may have been responsible for the acute myocardial infarction in our patient.

Angioplasty, Balloon, Coronary↗

PubMed1999

Influence of genotypes at SAA1 and SAA2 loci on the development and the length of latent period of secondary AA-amyloidosis in patients with rheumatoid arthritis.

To examine whether polymorphism at the SAA loci is associated with the development of amyloid protein A (AA)-amyloidosis, we determined the genotypes at the SAA1 and SAA2 loci in 43 AA-amyloidosis patients (amyloidosis population) and 77 patients with rheumatoid arthritis (RA) who had been ill for less than 5 years (early RA population). We also compared the frequencies of the genotypes at the SAA1 locus among 90 Korean, 95 Taiwanese, and 103 Japanese healthy subjects. The frequencies of the gamma/gamma genotype and gamma alleles at the SAA1 locus were significantly higher in the amyloidosis population than in the early RA population (34.9% versus 7.8%, and 58.1% versus 33.8%, chi2 test P=0.0001). The frequencies of the gamma allele at the SAA1 locus in Koreans, Taiwanese, and Japanese were 41.6%, 35.6%, and 37.4%, respectively. The length of the latent period of AA-amyloidosis was significantly longer in the patients with smaller numbers of the gamma allele at the SAA1 locus (Spearman's correlation coefficient: -0.42, P<0.05). On the other hand, the mean C-reactive protein (CRP) level during 2 years prior to the diagnosis of AA-amyloidosis was significantly higher in the patients with larger numbers of the gamma allele at the SAA1 locus (Spearman's correlation coefficient: 0.34, P<0.05). No significant association was found between amyloidosis and polymorphism at the SAA2 locus. We postulate that the allele SAA1gamma renders an RA patient susceptible to amyloidosis, possibly by affecting the severity of inflammation in RA.

Adult↗

PubMed1999

Up-regulation of low density lipoprotein receptor by a novel isobenzofranone derivative, MD-700.

Stimulatory effects of a novel isobenzofranone, MD-700, on low density lipoprotein (LDL) receptor activity were investigated in vitro and in vivo. MD-700 at 0.03 microg/ml elevated the expression of LDL receptor in HepG2 cells within 4 h. Corresponding to this, uptake of fluorescent labeled-LDL (3,3'-dioctadecylindocarbocyanine-LDL) by the cells increased linearly in time- and dose-dependent manner by MD-700 for up to 12 h. In the experiment using HepG2 cells transiently transfected with promoter-luciferase gene constructs, MD-700 increased luciferase activity in a dose-dependent manner from 0.03 to 0.1 microg/ml. In contrast, luciferase activity was not stimulated by MD-700 in construct with a deleted sterol regulatory element (SRE)-1, suggesting importance of SRE-1 in stimulation of the LDL receptor gene promoter by MD-700. Binding experiments on liver membranes from MD-700-treated hamsters showed about a 60% increase in 125I-labeled LDL binding. A Scatchard plot revealed that MD-700 increased the maximal binding without affecting binding affinity. In contrast to findings with an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, pravastatin, MD-700 had no effect on the sterol synthesis in hamster liver homogenates. These results suggest that MD-700 stimulates the expression of LDL receptor, presumably in a manner independent of change in sterol metabolism, and thereby promotes LDL clearance. Hypocholesterolemic actions of MD-700 in hamsters were then examined. MD-700 lowered serum cholesterol levels in hamsters fed normal chow or a high-fat diet. Fractionation of serum lipoproteins demonstrated that MD-700 selectively decreased LDL and very low density lipoprotein cholesterol. Dose-dependent decrease in serum cholesterol was also seen in hypercholesterolemic rats. Thus, the hypocholesterolemic action of MD-700 may be attributed to up-regulation of the LDL receptor, based on stimulation of the transcription of the LDL receptor gene. Although pravastatin stimulates LDL uptake and lowers serum cholesterol in a manner similar to that seen with MD-700, the mechanism responsible for hypocholesterolemic action appears to differ.

Animals↗

PubMed1999

Muscarinic receptor occupancy by biperiden in living human brain.

Anticholinergic drug is often used to treat extrapyramidal symptoms. We measured muscarinic cholinergic receptor (mAchR) occupancy by the oral administration of biperiden in eight healthy subjects using positron emission tomography (PET) and [11C]N-methyl-4-piperidylbenzilate (NMPB). After the baseline scan each subject underwent one or two post-dose PET scans. mAchR occupancy was 10-45% in the frontal cortex three hours after the oral administration of 4 mg of biperiden. The occupancy correlated with the plasma concentration of biperiden in a curvilinear manner.

Adult↗

PubMed1999

Human parathyroid hormone (1-34) increases urinary excretion of lysosomal enzymes in rats.

The kidney is the major target of parathyroid hormone (PTH), and PTH influences the urinary excretion of calcium, phosphate and hydrogen ions. It was previously reported that the urinary, excretion of N-acetyl-beta-D-glucosaminidase (NAG), a lysosomal enzyme, transiently increases after human PTH (hPTH) (1-34) infusion in normal subjects and idiopathic hypoparathyroidism patients, but not in pseudohypoparathyroidism type I patients. Here we report that intravenous infusion of hPTH(1-34) to rats transiently increased the urinary excretion of various lysosomal enzymes, such as beta-glucuronidase and acid phosphatase as well as NAG. However, it did not affect the urinary excretion of tubular brush border membrane enzymes, i.e. alkaline phosphatase, leucine aminopeptidase and gamma-glutamyl transpeptidase. Human PTH(1-34) dose-dependently increased the urinary excretion of NAG in rats with a peak at 30 min, which returned to a baseline within 60 min. The increase in the urinary NAG excretion caused by hPTH(1-34) positively correlated with the increase in the urinary cAMP excretion (r = 0.844, p < 0.01), and infusion of dibutyryl cAMP at a dose of 20 mg/kg similarly increased the urinary excretion of NAG. These results suggested that the increase in the urinary excretion of lysosomal enzymes caused by hPTH(1-34) may be a functional response to hPTH(1-34) occurring in the renal tubules via PTH signaling pathway.

Acetylglucosaminidase↗

PubMed1999

Determination of 99Tc deposited on the ground within the 30-km zone around the Chernobyl reactor and estimation of 99Tc released into atmosphere by the accident.

Technetium-99 was determined in samples from the 30-km zone around the Chernobyl reactor. Concentrations of 99Tc in soil samples taken from three forest sites ranged from 1.1 to 14.1 Bq kg-1 dry weight for the organic soil layers, and from 0.13 to 0.83 Bq kg-1 dry weight for the mineral soil layers. In particular, for the organic layers, the measured 99Tc concentrations were one or two orders of magnitude higher than those due to global fallout 99Tc. The 99Tc depositions (Bq m-2), based on the sum of the depositions measured in organic and mineral layers, ranged from 130 Bq m-2 within the 10-km zone to about 20 Bq m-2 close to the border of the 30-km zone. Taking the corresponding measured 137Cs depositions into account, it was found that the activity ratio of 99Tc/137Cs ranged from 6 x 10(-5) to 1.2 x 10(-4). It was estimated that about 970 GBq of 99Tc had been released by the Chernobyl accident. This figure corresponded to 2%-3% of the total 99Tc inventory in the core.

Air Pollutants, Radioactive↗